High-Resolution CGH Charaterization of Brain Tumors

脑肿瘤的高分辨率 CGH 表征

• 批准号: 6884670
• 负责人: SIMON G GREGORY
• 金额: $ 17.81万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-15 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6884670
• 关键词: artificial chromosomes; astrocytoma; biotechnology; brain disorder diagnosis; brain neoplasms; chromosome deletion; chromosome translocation; clinical research; comparative genomic hybridization; diagnosis design /evaluation; genetic mapping; genetic markers; glioblastoma multiforme; human tissue; microarray technology; neoplasm /cancer diagnosis; neoplasm /cancer genetics; neoplastic growth; nucleic acid amplification techniques; nucleic acid purification; oligodendroglia; oligonucleotides; polymerase chain reaction; prognosis; tissue /cell culture

DESCRIPTION (provided by applicant): It has been known for many years that malignant tumors have chromosome losses that contribute to their malignancy. In recent years, considerable research effort has been directed at identifying the one common region of deletion that occurs within specific tumor types in order to identify the actual gene(s) involved in the transformations. Traditionally, loss of heterozygosity mapping, using polymorphic markers and genotype information, and comparative genomic hybridization (CGH), utilizing normal metaphase chromosomes as the template upon which differentially labeled test and control samples are hybridized, have been used to identify chromosomal rearrangements. However, these approaches have proven to be relatively time consuming and have low resolution when identifying aberrant chromosomal regions. The project outlined here describes the development of high-resolution, chromosome specific, CGH arrays for the detection of chromosomal deletions and amplifications in neurological neoplasia. High-resolution array CGH will allow for very rapid and accurate (100 -200 kilobase) characterization of chromosomal rearrangements within a large number of tumors. High-resolution genome-wide coverage, on a per-chromosome basis, will be achieved by utilizing complete, overlapping, minimum-tile path clones that were generated by the Human Genome Project in its production of highly accurate genomic sequence. The research project contains four basic aims; aim 1, the generation of genome-wide 1Mb, chromosome and region specific high-resolution CGH arrays, including the purification of minimum-tile path DNA and production of amino linked DOP-PCR products; aim 2, the characterization of the arrays by testing with differentially labeled normal DNA's as well as known deletions and amplifications; aim 3, the characterization of clinically defined brain tumor types (glioblastoma multiforme, oligoastrocytoma and oligodendroglioma) by high-resolution CGH hybridization to determine exact regions of chromosomal rearrangement and therefore the putative disease causing genes within them; aim 4, investigate the use of high-resolution CGH arrays as a diagnostic tool for predicting the responsiveness of oligodendrogliomas to chemotherapeutic treatment that are associated with known chromosome deletions.

描述（由申请人提供）： 多年来，人们已经知道恶性肿瘤具有导致其恶性的染色体丢失。近年来，大量的研究工作已经针对确定在特定肿瘤类型中发生的一个常见的缺失区域，以确定参与转化的实际基因。传统上，杂合性缺失作图，使用多态性标记和基因型信息，和比较基因组杂交（CGH），利用正常中期染色体作为模板，差异标记的测试和对照样品进行杂交，已被用于识别染色体重排。然而，这些方法已被证明是相对耗时的，并且在鉴定异常染色体区域时具有低分辨率。 这里概述的项目描述了高分辨率，染色体特异性，CGH阵列的发展，用于检测染色体缺失和扩增的神经系统肿瘤。高分辨率阵列CGH将允许非常快速和准确的（100 - 200倍酶）表征大量肿瘤内的染色体重排。将通过利用人类基因组计划在生产高度准确的基因组序列时产生的完整、重叠、最小瓦片路径克隆，实现对每个染色体的高分辨率全基因组覆盖。 该研究项目包括四个基本目标：目标1，全基因组1 Mb、染色体和区域特异性高分辨率CGH阵列的产生，包括最小路径DNA的纯化和氨基连接的DOP-PCR产物的产生;目标2，通过与差异标记的正常DNA以及已知的缺失和扩增进行测试来表征阵列;目的3，临床定义的脑肿瘤类型的表征（多形性胶质母细胞瘤、少突星形细胞瘤和少突胶质细胞瘤）通过高分辨率CGH杂交以确定染色体重排的确切区域，从而确定其中的推定致病基因;目的4，研究使用高分辨率CGH阵列作为诊断工具，用于预测与已知染色体缺失相关的少突胶质细胞瘤对化疗治疗的反应性。